Senior of Histopathology Department at Khartoum, Radiation and Isotopes Center

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1 EUROPEAN ACADEMIC RESEARCH Vol. IV, Issue 2/ May 2016 ISSN Impact Factor: (UIF) DRJI Value: 5.9 (B+) Immune Histochemical Evaluation of AMACR (P504S) in Prostatic Adenocarcinoma and Benign Prostatic Hyperplasia in Small Neddle Biopsy of Sudanese EL TAYEB EL FAITH ELTAYEB MSc Student in Medical Laboratory Sciences AL-Neelain University, Sudan EL SADIG A ADAM Department of Patholothology Al-Ribat National University, Sudan NADA SALIH SALIH Senior of Histopathology Department at Khartoum, Radiation and Isotopes Center Abstract: Background: Prostate cancer is a major health problem throughout the world. Immunohistochemistry plays a very important role in the diagnosis of minimal prostatic adenocarcinoma and to exclude one of its benign mimickers, but it should always be interpreted in the context of the H&E appearances. In some cases of minimal prostate cancer morphologic features do not allow a diagnosis of carcinoma. Methods: 50 needle biopsy specimens, including 28 with small biopsy of prostatic adenocarcinoma and 22 benign prostate were stained immunohistochemically with AMACR. Results: Of 28 cases of small foci of prostatic adenocarcinoma, 26 (92.9%) expressed AMACR; There was focal positive staining with AMACR in 2 benign cases. 1380

2 Conclusions: Immunostaining with AMACR (p504s) could improve the diagnostic performance and help in avoid carrying out new biopsies in small foci of prostatic carcinoma detection. Key words: Prostate cancer, AMACR (P504S) marker, needle biopsy 1. INTRODUCTION Worldwide, prostate cancer is the second most common malignancy in men after lung cancer [1] Diagnosis of prostate cancer glands can sometimes present a diagnostic challenge for pathologists, since prostate carcinoma can mimic benign prostate glands [2]. The diagnosis of prostatic adenocarcinoma, especially in needle biopsy samples, can occasionally be challenging, either because they only show small foci of prostatic adenocarcinoma, or because of the difficulty in distinguishing prostatic carcinoma from benign mimickers[3] The difficulty in the diagnosis of prostatic adenocarcinoma is mostly seen with minimal (limited<1mm) carcinoma in needle tissue [4] Many major and minor histologic features important for the diagnosis of minimal prostatic carcinoma should be assessed specifically at low- and highpower magnification. The first of the major criteria is an infiltrative growth pattern which frequently presents as the presence of small malignant glands between larger, more complex (and often paler), benign glands. [5] Alpha-methylacyl- CoA racemase (AMACR), formerly known as P504s, is a mitochondrial and peroxisomal enzyme involved in the betaoxidation of branched fatty acids and bile acid intermediates [6] We aimed in this study to correlate between immunohistochemical expression of AMACR in small focal prostatic carcinoma and BPH in true cut needle biopsy. 1381

3 2. MATERIALS AND METHODS: 2.1. Materials: Subjects: (carcinoma group) with age range from 40 to 102 years (mean = 68.2) and Patient (BPH group) with age range from 60 to 85 years (mean = 72.5± 3.1) obtained from the department of pathology, Ibn sena hospital and Military hospital during the period from January 2015 to January 2016 were chosen for this study Samples: A total of 50 prostate needle biopsy specimens, including 28 prostate needle biopsy specimens with prostatic adenocarcinoma and 22 BPH The diagnosis of prostate cancer was established from: Examination of multiple levels of H&Estained sections and was confirmed by two pathologists Methods: Immunohistochemical Analysis Immunohistochemical staining was carried out using streptoavidin-biotin immunoperoxidase technique (thermo fisher). Three to five micrometer thick sections, cut from formalin fixed paraffin embedded blocks, were deparaffinized in Xylene and rehydrated in graded alcohol. The mounted sections were immersed in target retrieval solution, tris buffer EDTA (PH 9.0), then boiled in this solution in PT link for 20 min and then washed in phosphate buffer saline (ph 7.2).Then the slides were then incubated 20 minute using a polyclonal anti-amacr antibody ready to use thermofisher), After a buffer rinse, bound antibodies were detected with the thermo Envision System. Slides were counterstained with hematoxylin, and rinsed again. 1382

4 The slides were allowed to air dry and were cover slipped with permanent mounting media. Negative controls, in which the primary antibodies were replaced by PBS, were carried out for each primary antibody For AMACR, prostate carcinoma was used as positive internal control. - Immunohistochemical Evaluation: - Evaluation of AMACR: Results obtained from two sections were detected by the researchers and confirmed by experienced histopathologist. Negative and positive controls were used for evaluation of the test sections. Ethical clearance for this studies is provided by ethical committee of AL-Neelain University -faculty of medical laboratory science Statistical analysis The results of the study were statistically analyzed using SPSS version15 statistical program. Data were expressed as mean± SD for quantitative variables, numbers and percentage. For categorical variables, student t test was used. For statistical analysis of Gleason's grading Spearman's statistical test was used. P< 0.05 was considered the significant limit. 3. RESULTS: Staining results with AMACR AMACR expression in malignant glands had much more extensive and intensive staining results than benign glands (P<0.001). Prostatic carcinoma showed a brown cytoplasmic granular staining pattern of AMACR in the malignant glands and cells. A total of 50 prostate needle biopsy specimens The Frequency of adenocarcinoma in all sample are28 (56%)while The Frequency of benign prosatic hyperplasia is 22(44%) 1383

5 Table (1). Four specimens of prostatic adenocarcinomas were intermediate grade gleason (5-7) and twenty four were high grade Gleason (8-10). Out of 28 cases of small biopsy of prostatic carcinoma 26 (92.8%) expressed AMACR (p504s),and 2 did not express AMACR (p504s) while in twenty two cases of small biopsy of Benign prostatic hyperplasia have only 3(13.6%) expressed AMACR (p504s) Table(2). the expression of AMACR in prostatic adenocarcinoma is more than BPH which is statically significance ((p<0.0001)) Pvalue :0.00 Table (1) showing the frequency of prostatic adenocarcinoma and begin prostatic hyperplasia among study population Sample Frequency Percent Adenocarcinoma BPH Total Table (2):-showing the frequency of expression of AMACR among study population. Diagnosis AMCAR expression Total positive Negative Adenocarcinoma BPH Total DISCUSSION: In our finding we found that the expression of AMACR in prostate cancer (92%) is more than benign prostatic hyper plasia (9.9%) but some of benign are expressed. Our finding is in agreement with study done by Yang et al., 2002; Jiang et al., 2002 Beach et al Boran et al. 2011[7]; [8]; [9]; [10] which are found, 71% of cancer cases showed positive immune-staining with AMACR, but variable 1384

6 intensities and percentages of cells were present. About 71% 100% of prostatic adenocarcinoma stained with AMACR. and also this finding is inconsistent with other study. The sensitivity of AMACR in detecting these variants was found to be 70% by Farinola and Epstein 2004 [20], 68% by Zhou et al., 2003 [21] and 77% by Zhou et al., 2003) [21] respectively. 5. CONCLUSION Using AMACR as a positive marker alone might be misleading because weak expression of AMACR might be seen in benign glands. Accordingly, one should not render a diagnosis of benignancy based solely on a negative AMACR immune-stain. 6. ACKNOWLEDGEMENT: This work was supported by the work of Asma Siddig, Zuhal Ahmed Eltayeb, Nagla Mohammed Abu Elgasim and Rania Ahmed Hassan. 7. REFERENCES: 1- Ferlay J, Bray F, Pisani P. Globocan 2002: Cancer incidence, mortality and prevalence worldwide. Version 2.0. Lyon, France: IARC Press; IARC CancerBase No Gaudin PB, Reuter VE. Benign mimics of prostatic adenocarcinoma on needle biopsy. Anat Pathol 1997;2: Hameed O, Humphrey PA. Immunohistochemistry in the diagnosis of minimal prostate cancer Current Diagnostic Pathology 2006; 12: ) 4-Thorson P, Humphrey PA. Minimal adenocarcinoma in prostate needle biopsy tissue. Am J Clin Pathol 2000;114:

7 5-Epstein JI. Diagnostic criteria of limited adenocarcinoma of the prostate on needle biopsy. Hum Pathol. 1995; 26: Ferdinandusse S, Denis S, IJlst L, Dacremont G,Waterham HR, Wanders RJ.Subcellular localization and physiological role of alphamethylacyl- CoA racemase in humans. J Lipid Res 2000; 41: Yang XJ, Wu CL, Woda BA, et al. Expression of alphamethylacyl-coa racemase (P504S) in atypical adenomatous hyperplasia of the prostate. Am J Surg Pathol 2002; 26: ) 8-Jiang Z, Wu CL, Woda BA, et al. P504S/alphamethylacyl- CoA racemase: a useful marker for diagnosis of small foci of prostatic carcinoma on needle biopsy. Am J Surg Pathol 2002; 26: ) 9-Beach R, Gown AM, De Peralta-Venturina MN, et al. P504S immunohistochemical detection in 405 prostatic specimens including gauge needle biopsies. Am J Surg Pathol 2002; 26: Boran C, Kandirali E, Yilmaz F, Serin E, Akyol M. Reliability of the 34βE12, keratin 5/6, p63, bcl-2, and AMACR in the diagnosis of prostate carcinoma. Urologic Oncology: Seminars and Original Investigations 2011; 29: Farinola MA, Epstein JI. Utility of immunohistochemistry for alpha-methylacyl- CoA racemase in distinguishing atrophic prostate cancer from benign atrophy. Hum Pathol 2004; 35: Zhou M, Jiang Z, Epstein JI. Expression and diagnostic utility of alpha-methylacyl-coaracemase (P504S) in foamy gland and pseudohyperplastic prostate cancer. Am J Surg Pathol 2003; 27:

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